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Competitive SARS-CoV-2 Serology Reveals Most Antibodies Targeting the Spike Receptor-Binding Domain Compete for ACE2 Binding.
mSphere. 2020 09 16; 5(5)M

Abstract

As severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread around the world, there is an urgent need for new assay formats to characterize the humoral response to infection. Here, we present an efficient, competitive serological assay that can simultaneously determine an individual's seroreactivity against the SARS-CoV-2 Spike protein and determine the proportion of anti-Spike antibodies that block interaction with the human angiotensin-converting enzyme 2 (ACE2) required for viral entry. In this approach based on the use of enzyme-linked immunosorbent assays (ELISA), we present natively folded viral Spike protein receptor-binding domain (RBD)-containing antigens via avidin-biotin interactions. Sera are then competed with soluble ACE2-Fc, or with a higher-affinity variant thereof, to determine the proportion of ACE2 blocking anti-RBD antibodies. Assessment of sera from 144 SARS-CoV-2 patients ultimately revealed that a remarkably consistent and high proportion of antibodies in the anti-RBD pool targeted the epitope responsible for ACE2 engagement (83% ± 11%; 50% to 107% signal inhibition in our largest cohort), further underscoring the importance of tailoring vaccines to promote the development of such antibodies.IMPORTANCE With the emergence and continued spread of the SARS-CoV-2 virus, and of the associated disease, coronavirus disease 2019 (COVID-19), there is an urgent need for improved understanding of how the body mounts an immune response to the virus. Here, we developed a competitive SARS-CoV-2 serological assay that can simultaneously determine whether an individual has developed antibodies against the SARS-CoV-2 Spike protein receptor-binding domain (RBD) and measure the proportion of these antibodies that block interaction with the human angiotensin-converting enzyme 2 (ACE2) required for viral entry. Using this assay and 144 SARS-CoV-2 patient serum samples, we found that a majority of anti-RBD antibodies compete for ACE2 binding. These results not only highlight the need to design vaccines to generate such blocking antibodies but also demonstrate the utility of this assay to rapidly screen patient sera for potentially neutralizing antibodies.

Authors+Show Affiliations

Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.Weill Institute for Neurosciences, University of California, San Francisco, San Francisco, California, USA. Department of Neurology, University of California, San Francisco, San Francisco, California, USA.Department of Laboratory Medicine, University of California, San Francisco, San Francisco, California, USA. Department of Medicine, University of California, San Francisco, San Francisco, California, USA.Chan Zuckerberg Biohub, San Francisco, California, USA. Department of Medicine, Stanford University Medical School, Stanford, California, USA. Department of Microbiology and Immunology, Stanford University Medical School, Stanford, California, USA.Weill Institute for Neurosciences, University of California, San Francisco, San Francisco, California, USA. Department of Neurology, University of California, San Francisco, San Francisco, California, USA.Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA jim.wells@ucsf.edu. Chan Zuckerberg Biohub, San Francisco, California, USA. Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, California, USA.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

32938700

Citation

Byrnes, James R., et al. "Competitive SARS-CoV-2 Serology Reveals Most Antibodies Targeting the Spike Receptor-Binding Domain Compete for ACE2 Binding." MSphere, vol. 5, no. 5, 2020.
Byrnes JR, Zhou XX, Lui I, et al. Competitive SARS-CoV-2 Serology Reveals Most Antibodies Targeting the Spike Receptor-Binding Domain Compete for ACE2 Binding. mSphere. 2020;5(5).
Byrnes, J. R., Zhou, X. X., Lui, I., Elledge, S. K., Glasgow, J. E., Lim, S. A., Loudermilk, R. P., Chiu, C. Y., Wang, T. T., Wilson, M. R., Leung, K. K., & Wells, J. A. (2020). Competitive SARS-CoV-2 Serology Reveals Most Antibodies Targeting the Spike Receptor-Binding Domain Compete for ACE2 Binding. MSphere, 5(5). https://doi.org/10.1128/mSphere.00802-20
Byrnes JR, et al. Competitive SARS-CoV-2 Serology Reveals Most Antibodies Targeting the Spike Receptor-Binding Domain Compete for ACE2 Binding. mSphere. 2020 09 16;5(5) PubMed PMID: 32938700.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Competitive SARS-CoV-2 Serology Reveals Most Antibodies Targeting the Spike Receptor-Binding Domain Compete for ACE2 Binding. AU - Byrnes,James R, AU - Zhou,Xin X, AU - Lui,Irene, AU - Elledge,Susanna K, AU - Glasgow,Jeff E, AU - Lim,Shion A, AU - Loudermilk,Rita P, AU - Chiu,Charles Y, AU - Wang,Taia T, AU - Wilson,Michael R, AU - Leung,Kevin K, AU - Wells,James A, Y1 - 2020/09/16/ PY - 2020/9/17/entrez PY - 2020/9/18/pubmed PY - 2020/9/29/medline KW - COVID-19 KW - SARS-CoV-2 KW - angiotensin-converting enzyme 2 KW - immunoserology KW - neutralizing antibodies KW - receptor-binding domain KW - serology JF - mSphere JO - mSphere VL - 5 IS - 5 N2 - As severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread around the world, there is an urgent need for new assay formats to characterize the humoral response to infection. Here, we present an efficient, competitive serological assay that can simultaneously determine an individual's seroreactivity against the SARS-CoV-2 Spike protein and determine the proportion of anti-Spike antibodies that block interaction with the human angiotensin-converting enzyme 2 (ACE2) required for viral entry. In this approach based on the use of enzyme-linked immunosorbent assays (ELISA), we present natively folded viral Spike protein receptor-binding domain (RBD)-containing antigens via avidin-biotin interactions. Sera are then competed with soluble ACE2-Fc, or with a higher-affinity variant thereof, to determine the proportion of ACE2 blocking anti-RBD antibodies. Assessment of sera from 144 SARS-CoV-2 patients ultimately revealed that a remarkably consistent and high proportion of antibodies in the anti-RBD pool targeted the epitope responsible for ACE2 engagement (83% ± 11%; 50% to 107% signal inhibition in our largest cohort), further underscoring the importance of tailoring vaccines to promote the development of such antibodies.IMPORTANCE With the emergence and continued spread of the SARS-CoV-2 virus, and of the associated disease, coronavirus disease 2019 (COVID-19), there is an urgent need for improved understanding of how the body mounts an immune response to the virus. Here, we developed a competitive SARS-CoV-2 serological assay that can simultaneously determine whether an individual has developed antibodies against the SARS-CoV-2 Spike protein receptor-binding domain (RBD) and measure the proportion of these antibodies that block interaction with the human angiotensin-converting enzyme 2 (ACE2) required for viral entry. Using this assay and 144 SARS-CoV-2 patient serum samples, we found that a majority of anti-RBD antibodies compete for ACE2 binding. These results not only highlight the need to design vaccines to generate such blocking antibodies but also demonstrate the utility of this assay to rapidly screen patient sera for potentially neutralizing antibodies. SN - 2379-5042 UR - https://www.unboundmedicine.com/medline/citation/32938700/Competitive_SARS_CoV_2_Serology_Reveals_Most_Antibodies_Targeting_the_Spike_Receptor_Binding_Domain_Compete_for_ACE2_Binding_ L2 - https://doi.org/10.1128/mSphere.00802-20 DB - PRIME DP - Unbound Medicine ER -